• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.33-39
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• 2002

This study was conducted to investigate the effect of $Ca^{2+}$ concentration in fusion medium on the fusion, nuclear morphology and the development of bovine somatic cell nuclear transfer embryos. Bovine skin cells were transferred into an enucleated oocyte and fused with cytoplasm in the fusion medium containing with 0.05 to 1.0 mM Cacl$_2$. Nuclear transfer embryos were activated with a combination of A23187 and cycloheximide. Nuclear transfer embryos were fixed at 3 h after fusion or cultured for 7 ~8 days. Fusion rate was significantly (P＜0.01) increased by increasing the $Ca^{2+}$ concentrations in the fusion medium from 0.05 mM (56.6%) to 0.5 mM (50.1%) and 1.0 mM (84.3%). More than 80% of reconstituted embryos underwent premature chromosome condensation (PCC) with 0.05, 0.1 mM CaCl$_2$, whereas 54.5% and 59.3% of embryos formed pronucleus (PN) directly without PCC in the 0.5 and 1.0 mM CaCl$_2$, groups. Blastocyst formation rates were significantly (P＜0.05) different between 0.1 mM and 1.0 mM CaCl$_2$groups. From the present result, it is suggested that the elevated $Ca^{2+}$ concentrations in fusion medium can enhance the fusion and blastocyst formation rates of bovine nuclear transfer embryos.bryos.

• The Korean Journal of Pharmacology
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• v.32 no.2
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• pp.221-231
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• 1996

The properties of cytosolic $Ca^{2+}$ level$([Ca^{2+}]_i)$ movement of high KCl, carbachol and oxytocin were examined with myometrium isolated from non-pregnant rat(estrus cycle). High concentration of KCl$({\leq}23.3mM)$ induced rhythmic increases in $[Ca^{2+}]_i$ and muscle contraction. However, sustained $[Ca^{2+}]_i$ and contracion were obtained at higher KCl concentration $({\geq}30.3mM)$ The rhythmic and sustained contraction closely associated with changes in $[Ca^{2+}]_i$ induced by high KCl. Carbachol $(3{\sim}30{\mu}M$ generated rhythmic increases with tonic component in $[Ca^{2+}]_i$ and muscle contraction. Myometrial contraction stimulated by carbachol was also closely correlated with change in $[Ca^{2+}]_i$. And the $[Ca^{2+}]_i/contraction$ relationships were similar when muscle strips were stimulated by high KCl and carbachol. Maximal concentration of carbachol $(10{\mu}M)$ and oxytocin(100 nM) increased $[Ca^{2+}]_i$ and contraction which were slightly greater than that of high KCl in non-pregnant myometrium, respectively. However, the $[Ca^{2+}]_i$ and contraction were strongly inhibited by verapamil $(10{\mu}M)$, a 1-type $Ca^{2+}$ channel blocker, as in the case of high KCl. Additionally, although carbachol further increased $[Ca^{2+}]_i$ and contraction induced by high KCl, these changes also strongly inhibited by application of verapamil. These results suggest that uterotonic agents, carbachol and oxytocin, induced contraction by increase in $[Ca^{2+}]_i$ through $Ca^{2+}$ influx than by a regulation of $Ca^{2+}-sensitization$ in non-pregnant myometrium.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.2
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• pp.105-111
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• 2010

Inositol 1,4,5-trisphosphate receptors ($InsP_3Rs$) modulate $Ca^{2+}$ release from intracellular $Ca^{2+}$ store and are extensively expressed in the membrane of endoplasmic/sarcoplasmic reticulum and Golgi. Although caffeine and 2-aminoethoxydiphenyl borate (2-APB) have been widely used to block $InsP_3Rs$, the use of these is limited due to their multiple actions. In the present study, we examined and compared the ability of caffeine and 2-APB as a blocker of $Ca^{2+}$ release from intracellular $Ca^{2+}$ stores and $Ca^{2+}$ entry through store-operated $Ca^{2+}$ (SOC) channel in the mouse pancreatic acinar cell. Caffeine did not block the $Ca^{2+}$ entry, but significantly inhibited carbamylcholine (CCh)-induced $Ca^{2+}$ release. In contrast, 2-APB did not block CCh-induced $Ca^{2+}$ release, but remarkably blocked SOC-mediated $Ca^{2+}$ entry at lower concentrations. In permeabilized acinar cell, caffeine had an inhibitory effect on InsP3-induced $Ca^{2+}$ release, but 2-APB at lower concentration, which effectively blocked $Ca^{2+}$ entry, had no inhibitory action. At higher concentrations, 2-APB has multiple paradoxical effects including inhibition of Ins$P_3$-induced $Ca^{2+}$ release and direct stimulation of $Ca^{2+}$ release. Based on the results, we concluded that caffeine is useful as an inhibitor of $InsP_3R$, and 2-APB at lower concentration is considered a blocker of $Ca^{2+}$ entry through SOC channels in the pancreatic acinar cell.

• The Korean Journal of Physiology
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• v.20 no.1
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• pp.31-41
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• 1986

The sufficient myoplasmic $Ca^{++}$ to react with the contractile proteins is necessary to induce contraction of a cardiac muscle. These $Ca^{++}$ for the production of muscle contraction are supplied from the three recognized $Ca^{++}$ sources; internal $Ca^{++}$ release via the sarcoplasmic reticulum(SR), $Ca^{++}$ influx through a gated Ca-channel in the membrane as a Isi, and $Ca^{++}$ transport by the mechanism of Na/ca exchange. However, it is still controversial which $Ca^{++}$ sources act as a main contributor for myoplasmic $Ca^{++}$, Therefore, this study was undertaken in order to examine the $Ca^{++}$ sources for the contraction of frog ventricle. There is evidence that the SR is sparse in frog ventricular fibers, and that T-tubules are absent. Isolated ventricular strips of frog, Rana nigromaculata, were used in this experiment. Isometric tension was recorded by force transducer, and membrane potentials of ventricular muscles were measured through the intracellular glass microelectrodes, which were filled with 3M KCI and had resistance of $30{\pm}50M{\Omega}$. All experiments were performed at room temperature in a tris·buffered Ringer solution which was aerated with 100% $O_2$. Isotonic high K, low Na solution was used to induce K-contracture, K-contracture appeared at the concentration of 20 to 30mM-KCI and was potentiated in parallel with the increase in KCI concentration. The contracture had two components: an initial rapid phasic and a subsequent slow tonic contractile responses. Membrane Potentials measured at normal Ringer solution(2.5mM KCI) was -90 to -100 mV, and decreased linearly as the KCI concentration increased; -55mV at 20mM.KCI, -45mV at 30 mM.KCI, -30 mY at 50 mM.KCI, and -12 mV at 100 mM.KCI. K-contracture was evoked firstly at the membrane potential of -45 mV. The contracture was potentiated by the increase of bathing extracellular $Ca^{++}$ concentration. However, in the absence of $Ca^{++}$ the contracture was almost not induced by 50 mM.KCI solution. Caffeine(20mM) in normal Ringer solution, which is known to release $Ca^{++}$ from SR without substantial effects on the $Ca^{++}$ fluxes across the surface membrane, did not affect membrane potential and also not initiate contracture, but the caffeine in 20 mM-KCI Ringer solution produced a contracture. Above results suggest that the main $Ca^{++}$ source for the K·contracture of frog ventricle is $Ca^{++}$ influx through the voltage-dependent Ca-channel, and that in the K-contracture at the concentration of 100 mM-KCI, the mechanism of Na/ca exchange also partly contributs, in addition to the $Ca^{++}$ influx.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.373-380
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• 1993

When the cells of yeast K35 were immobilized in Ca-alginate gel, cell concentration and viability decreased as alginate concentration increased. Considering the results, 2% (w/v) Ca-alginate concentration would be suitable. Among various concentrations of additives and cross-lin-king agent, the addition of 1.67% (w/v) of bentonite together with 0.33% (v/v) of glutaraldehyde (ABG bead) resulted in the highest ethanol production of 1.8%(w/v), using YPD medium containing 2% glucose. ABG bead seemed to be more resistant to phosphate ion than Ca-alginate bead. 0.33%(w/v) of phosphate was a proper concentration for the ethanol production by ABG bead. Scanning electron microscopic observation depicted that the immobilized cells on the bead surface were coated by alginate gel and that the cells in the internal bead were cross-linked with alginate matrix. When repeated-batch culture was performed with ABG bead for 40 days in a packed-bed reactor, ethanol concentration of about 90~110 g/l-gel was maintained. Cell viability was maintained around 70%, and outgrowing cell concentration was below 6.3% of total cell concentration. Consequently, the results showed that ABG head was a potential carrier for continuous production of ethanol compared to conventional Ca-alginate bead.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.3
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• pp.219-227
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• 2019

Polycystic kidney disease 2-like-1 (PKD2L1), polycystin-L or transient receptor potential polycystin 3 (TRPP3) is a TRP superfamily member. It is a calcium-permeable non-selective cation channel that regulates intracellular calcium concentration and thereby calcium signaling. Although the calmodulin (CaM) inhibitor, calmidazolium, is an activator of the PKD2L1 channel, the activating mechanism remains unclear. The purpose of this study is to clarify whether CaM takes part in the regulation of the PKD2L1 channel, and if so, how. With patch clamp techniques, we observed the current amplitudes of PKD2L1 significantly reduced when co-expressed with CaM and $CaM{\triangle}N$. This result suggests that the N-lobe of CaM carries a more crucial role in regulating PKD2L1 and guides us into our next question on the different functions of two lobes of CaM. We also identified the predicted CaM binding site, and generated deletion and truncation mutants. The mutants showed significant reduction in currents losing PKD2L1 current-voltage curve, suggesting that the C-terminal region from 590 to 600 is crucial for maintaining the functionality of the PKD2L1 channel. With PKD2L1608Stop mutant showing increased current amplitudes, we further examined the functional importance of EF-hand domain. Along with co-expression of CaM, ${\triangle}EF$-hand mutant also showed significant changes in current amplitudes and potentiation time. Our findings suggest that there is a constitutive inhibition of EF-hand and binding of CaM C-lobe on the channel in low calcium concentration. At higher calcium concentration, calcium ions occupy the N-lobe as well as the EF-hand domain, allowing the two to compete to bind to the channel.

• YAKHAK HOEJI
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• v.38 no.1
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• pp.57-66
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• 1994

We studied the effect of eicosanoids on the content of energy metabolites and the intrasynaptosomal $Ca^{++}([Ca^{++}]_i)$ concentration in cerebral ischemic rats. An ischemic model was made by bilateral carotid artery ligation (BCAL) and by incubation of synaptosomes under aglycemic and $N_2$ gas bubbling condition. The content of ATP, creatine phosphate and glucose decreased at 15 minutes after BCAL while that of lactate increased in male Wistar rats. Oral administration of EPA(100 mg/ml/Kg/day) or DHA(16 mg/ml/Kg/day) for 6 weeks improved both the decreases and the increase of the cerebral energy metabolites. In addition, the increase of $[Ca^{++}]_i$, under BACL was suppressed by EPA or DHA treatment. When the both Wistar rats and SHR were administered orally with EPA or DHA for 6 weeks, the effect on the increase of $[Ca^{++}]_i$ under ischemia by $N_2$ gas bubbling were protected. From these results, it may be that EPA or DHA treatment were greatly contributed to preservation of ischemic cerebral energy metabolism and $Ca^{++}$ concentration.

• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.81-81
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• 2003

Glibenclamide, a sulfonylurea derivative, has been used in tile treatment of type II diabetes mellitus. Recent studies provided evidence that glibenclamide, in addition to blocking ATP-sensitive $K^{+}$ channels, also affected Na$^{+}$-K$^{+}$ pumps and L-type $Ca^{2+}$ channels in noncardiac cells. The effect of glibenclamide on the cardiac muscle is not clearly known. In the present study, the effects of glibenclamide on intracellular Na$^{+}$ concentration ([Na$^{+}$]$_{i}$ ), twitch tension, $Ca^{2+}$ transient, and membrane potential were investigated in isolated guinea-pig ventricular myocytes. Glibenclamide at concentration of 200 $\mu$M increased [Na$^{+}$]$_{i}$ by 3.9$\pm$0.4 mM (mean $\pm$ SE, n=12), decreased twitch tension by 36.1 $\pm$ 4.0％ (mean $\pm$ SE, n=8), reduced $Ca^{2+}$ transient by 24.4$\pm$5.1％ (mean $\pm$ SE, n=3), slightly depolarized diastolic membrane potential, and did not change action potential duration. To determine whether inhibitions of Na$^{+}$-K$^{+}$ pumps and L-type $Ca^{2+}$ channels are responsible for the increase of [Na$^{+}$]$_{i}$ and the decrease of twitch tension, we tested effects of glibenclamide on Na$^{+}$-K$^{+}$ pump current and L-type $Ca^{2+}$ current. Glibenclamide decreased Na$^{+}$-K$^{+}$ pump current and L-type $Ca^{2+}$ current in a concentration-dependent manner.t in a concentration-dependent manner.

• Clean Technology
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• v.16 no.2
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• pp.124-131
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• 2010

Effects of $Ca(OH)_2$ concentration (0.16~0.64 wt%), surfactant concentration (2~16 wt%), total volumetric flow rate (3~6 L/min) and $CO_2$ volume fraction $(0.3{\sim}0.6)$ on morphology, crystal structure, mean particle diameter, aggregation and specific surface area of the precipitated $CaCO_3$ were investigated in the slurry bubble column reactor. Experiments were carried out in acrylic reactor ($0.11\;m-ID{\times}1.0\;m-high$) with a internal tube ($0.04\;m-ID{\times}1.0\;m-high$h). The reaction time of $CaCO_3$ synthesis decreased with adding Dispex N40 of the anionic surfactant. The reaction rate of $Ca(OH)_2$ increased with increasing the volumetric flow rate of $CO_2$. From SEM images, the single crystal of $CaCO_3$ increased with increasing the reaction rate in the saturated concentration of $Ca(OH)_2$ (0.16 wt %) and the concentration of Dispex N40 (2 wt%). The mean particle size of $CaCO_3$ varied with adding Dispex N40. In addition, the specific surface area of $CaCO_3$ increased with adding of surfactant (2 wt%) from $35m^2/g$ to $44m^2/g$ at the volumetric flow rate of $CO_2$ (0.9 L/min) and the concentration of $Ca(OH)_2$(0.64 wt %).

• The Korean Journal of Physiology
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• v.19 no.1
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• pp.1-13
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• 1985

The influences of $Ca^{2+}-antagonists$, verapamil and $Mn^{2+}$, upon the spontaneous electrical activity and contractions were studied in guinea-pig taenia coli. Spontaneous contractions were recorded with force transducer, and spike action potentials were measured extracellularly by use of suction electrode. All experiments were performed in tris-buffered Tyrode solution Which was aerated With 100% $O_2$ and kept at $35^{\circ}C$. The results obtained were as follows : 1) Verapamil suppressed the frequency and amplitude of spontaneous contractions dose dependently, and blocked completely mechanical responses at the concentration of 1 mg/1. 2) The frequency of bursts of spike discharge(bursts frequency) and the number of spikes in a burst(spikes frequency) were reduced in a dose·dependent manner within the concentration range of $10^{-5}$ to $10^{-3}g/l$, and bursts frequency was affected more readily at a low concentration of $10^{-5}g/l$ verapamil. 3) The verapamil_induced suppression of spontaneous contractions in the Tyrode solution containing 1 mM $Ca^{2+}$ was completely antagonized by the addition of extra $Ca^{2+}$ to the Tyrode solution $(2\;m\;MCa^{2+})$. 4) $Mn^{2+}$ suppressed the amplitude of spontaneous contractions, whereas $Mn^{2+}$ accelerated the frequency dose-dependently within the range of low concentrations$(10^{-7}\;to\;10^{-4}\;M\;Mn^{2+})$. 5) The bursts frequency determining frequency of spontaneous contractions was increased in a dose-dependent manner, whereas the spikes frequency known to determine the contractions amplitude was reduced within the range of low concentrations. At a high concentration of 1 mM $Mn^{2+}$, however, all spontaneous contractile responses were blocked simultaneously with the disapperance of electrical activity. 6) The frequency and amplitude of spontaneous contractions altered by $Mn^{2+}$ in 1mM $Ca^{2+}$ Tyrode were increased by extra $Ca^{2+}(2mM)$.